JP2014055841A - Microchip and analyzing device using microchip - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a microchip that has a simple and compact structure, and makes it possible to simply conduct a desired analysis, and an analyzing device using the microchip.SOLUTION: A microchip 10 includes a chamber 1 in which particles are stored and the particles are irradiated with light, and a storage part 2 that is disposed in a channel 5 for making a liquid sample flow into the chamber 1, and has particles in a dried state stored therein, and the microchip 10 is used to detect the agglomeration of particles.

Description

  The present invention relates to a microchip and an analyzer using the microchip.

  In recent years, the importance of detecting and quantifying biological substances (for example, nucleic acids, proteins, cells, viruses, etc.) or chemical substances in various fields such as medical care, health, food, and drug discovery has increased. Attention has been focused on the development of microchips for the detection and quantification of urine.

  For example, Patent Document 1 discloses a microchip intended to improve the accuracy of analysis. Specifically, Patent Document 1 discloses a liquid reagent holding unit containing a liquid reagent used for analysis, a plurality of mixing units for mixing the liquid reagent and a sample to be analyzed, and the above mixing. There is disclosed a microchip having a residue separation part for separating the resulting residue.

JP 2009-281869 A (released on December 3, 2009)

  However, when the reagent is stored in a liquid state as described above, there is a problem that the activity of the reagent such as particles tends to decrease. And when the activity of a reagent falls, the problem that the value detected using the said reagent will become a value different from the original value generate | occur | produces.

  The present invention has been made in view of the above-described conventional problems, and an object of the present invention is to provide a microchip that has a simple and small structure and can easily perform a desired analysis, and the microchip. It is to provide an analysis apparatus used.

  In order to solve the above problems, the microchip of the present invention is a microchip for detecting the aggregation of particles. The microchip accommodates the particles and is irradiated with light for the detection. And a storage unit that is disposed in a flow path for flowing a liquid sample into the chamber and stores the particles in a dry state.

  According to the above configuration, the storage unit is arranged in the flow path for flowing the liquid sample into the chamber, and the dried particles are stored in the storage unit. Therefore, when a liquid sample is caused to flow into the flow path, the liquid sample reaches the storage unit, and particles are dissolved in the liquid sample that has reached the storage unit.

  Thereafter, the particles dissolved in the liquid sample are accommodated in the chamber while being in contact with the liquid sample. The particles that are in contact with the liquid sample aggregate depending on the components contained in the liquid sample. In addition, the movement of the particles from the storage unit to the chamber is performed by the flow of the liquid sample in the flow path.

  In the chamber, the particles are irradiated with light, and the aggregation of the particles is detected by detecting the resulting transmitted light or scattered light (forward scattered light and / or back scattered light).

  That is, according to the above configuration, by one operation of flowing the liquid sample into the flow path, the particles are dissolved in the liquid sample, the dissolved particles are moved to the chamber, the light in the chamber is irradiated with light, and transmitted. A plurality of steps of detecting light or scattered light can be performed continuously.

  In the microchip of the present invention, the storage part is preferably formed of a porous substrate.

  According to the said structure, particle | grains can be preserve | saved with high density in the hole of a porous base material. Moreover, according to the said structure, since the contact area of particle | grains and a liquid sample can be enlarged easily, particle | grains can be efficiently dissolved in a liquid sample.

  In the microchip of the present invention, a diffusion unit including a structure for dispersing the particles moving in the flow channel is provided at a position between the storage unit and the chamber in the flow channel. Is preferred.

  The particles dissolved in the liquid sample in the storage unit move in the flow path while causing streamline density unevenness. Since the diffusion unit is provided on the downstream side of the storage unit, the particles dissolved in the liquid sample in the storage unit reach the diffusion unit. And since the spreading | diffusion part is equipped with the structure which disperse | distributes a particle | grain, the particle | grains which move the inside of a flow path can be disperse | distributed uniformly (in other words, a density nonuniformity can be eased).

  Since the uniformly dispersed particles are accommodated in the chamber in a uniformly dispersed state, the density of the particles in the chamber becomes uniform. At this time, even if any portion of the chamber is irradiated with light, the particles in the same dispersed state are irradiated with light.

  Therefore, according to the above configuration, the aggregation of particles can be detected with high accuracy without being influenced by the detection location in the chamber.

  In the microchip of the present invention, the structure is preferably a plurality of pillar-shaped structures.

  According to the said structure, the particle | grains which move the inside of a flow path can be disperse | distributed more uniformly (in other words, a density nonuniformity can be eased more). As a result, the accuracy of the particle aggregation detection result can be further increased.

  In the microchip of the present invention, the structure is preferably a porous substrate.

  According to the said structure, the particle | grains which move the inside of a flow path can be disperse | distributed more uniformly (in other words, a density nonuniformity can be eased more). As a result, the accuracy of the particle aggregation detection result can be further increased.

  In the microchip of the present invention, the channel is preferably provided with an injection part for injecting a liquid sample into the channel.

  According to the said structure, a liquid sample can be easily inject | poured in a flow path through an injection | pouring part.

  In the microchip of the present invention, the particles are preferably latex particles or latex particles to which an antibody is bound.

  According to the said structure, the microchip of this invention can be used as a microchip (microchip for latex aggregation immunoturbidimetry) for implementing a latex agglutination immunoturbidimetry.

  In the microchip of the present invention, the chamber includes at least a first substrate having light transmittance to which the light is irradiated, and at least a part of light incident on the chamber through the first substrate. And a second substrate that reflects toward the inside.

  According to the above configuration, light entering the chamber through the first substrate is irradiated to the particles in the chamber in the process of traveling from the first substrate toward the second substrate. At this time, backscattered light is generated from the particles toward the first substrate. Further, according to the above configuration, at least a part of the light incident into the chamber through the first substrate is reflected to the inside of the chamber by the second substrate, and the reflected light is irradiated to the particles in the chamber. Is done. At this time, forward scattered light is generated from the particles toward the first substrate. That is, according to the said structure, backscattered light and forward scattered light can be generated toward the same side. Then, by simultaneously measuring the back scattered light and the forward scattered light (for example, measured by one detection unit), the detection sensitivity and detection accuracy of the scattered light can be dramatically improved.

  In order to solve the above-mentioned problems, the analyzer of the present invention has a light source for irradiating light to the chamber of the microchip of the present invention, and a side opposite to the light source on the detection surface in the chamber. And a detector for detecting transmitted light after passing through the chamber, which is provided on an extension line in the irradiation direction of light emitted from the light source.

  According to the above configuration, since the transmitted light can be efficiently detected by the detection unit, the aggregation of particles can be detected with high accuracy based on the transmitted light.

  In order to solve the above-described problems, the analyzer of the present invention has a light source for irradiating light to the chamber of the microchip of the present invention, and a detection surface in the chamber on the same side as the light source. And a detection unit for detecting scattered light, which is provided.

  According to the above configuration, since the scattered light can be detected efficiently by the detection unit, the aggregation of particles can be detected with high accuracy based on the scattered light.

  In the analyzer of the present invention, it is preferable that at least one of an optical slit and a condenser lens is provided in the optical path between the light source and the microchip.

  According to the above configuration, the particles in the chamber can be efficiently irradiated with light (in other words, stray light can be suppressed), so that detection sensitivity and detection accuracy of transmitted light or scattered light are improved. Can be made. As a result, the aggregation of particles can be detected with high accuracy.

  In the analyzer according to the present invention, the light source may include an irradiation area of light incident on the chamber in a direction perpendicular to the liquid feeding direction in the chamber, of the wall surface surrounding the internal space of the chamber. The first wall surface that makes light incident on the first wall surface and the second wall surface that is irradiated after the light incident from the first wall surface travels straight through the chamber may not overlap with the other wall surfaces. preferable.

  When the wall surface is irradiated with light, the light is scattered by the wall surface. As a result, when scattered light is detected, the background noise increases, and when transmitted light is detected, the amount of light detected is reduced, so the transmittance is estimated to be higher than actual. Problem arises. However, according to the said structure, the problem mentioned above can be eliminated by minimizing the scattering of the light by a wall surface.

  In the analyzer of the present invention, the light source has a length in a direction perpendicular to the liquid feeding direction in the chamber of an irradiation area of light incident on the chamber in a direction perpendicular to the liquid feeding direction in the chamber. It is preferable that the thickness can be adjusted.

  According to the above configuration, even if the particles accommodated in the chamber have uneven density, the aggregation state of the particles can be detected by averaging the length of the light irradiation area. . That is, according to the above configuration, the aggregation of particles can be detected with higher accuracy.

  For example, as the length of the light irradiation area is adjusted to be longer, the aggregation state of the particles can be detected by averaging more.

  In the analyzer of the present invention, the light source has a length in a direction perpendicular to the liquid feeding direction in the chamber of an irradiation area of light incident on the chamber in a direction perpendicular to the liquid feeding direction in the chamber. It is preferable to adjust the length to 80% or more of the length of the chamber in a direction perpendicular to the liquid feeding direction of the chamber.

  According to the above configuration, it is possible to more reliably average and detect the aggregation state of the particles.

  In the analyzer of the present invention, it is preferable that the light irradiation direction of the light source is parallel to the liquid feeding direction in the chamber in a direction horizontal to the detection surface in the chamber.

  When light is irradiated on the wall surface of the chamber, the light is scattered by the wall surface. As a result, when scattered light is detected, the background noise increases, and when transmitted light is detected, the amount of light detected is reduced, so the transmittance is estimated to be higher than actual. Problem arises. However, according to the said structure, the problem mentioned above can be eliminated by minimizing the scattering of the light by a wall surface.

  According to the present invention, since the activity of a reagent such as a particle can be retained in the chip, it can be accurately determined using the chip.

  According to the present invention, since the dried particles are used, the microchip can be stored for a long period of time without degrading the function of the microchip.

(A) And (b) is a figure which shows one Embodiment of the microchip of this invention. (A) And (b) is a figure which shows one Embodiment of the microchip of this invention. It is a figure which shows one Embodiment of the chamber in the microchip of this invention. It is a figure which shows one Embodiment of the chamber in the microchip of this invention. (A) And (b) is a figure which shows one Embodiment of the analyzer of this invention. It is a figure which shows one Embodiment of the optical slit in the analyzer of this invention. It is a figure which shows one Embodiment of the light source in the analyzer of this invention. It is a figure which shows one Embodiment of the analyzer of this invention. (A) And (b) is a figure which shows the manufacturing process of the microchip in the Example of this invention, and the outline of a structure of a microchip. It is a figure which shows the outline of a structure of the microchip in the Example of this invention. It is a figure which shows the outline of a structure of the microchip in the Example of this invention. It is a graph which shows the result of the analytical test using the microchip in the Example of this invention.

  An embodiment of the present invention will be described below, but the present invention is not limited to this. The present invention can be modified in various ways within the scope of the claims, and the present invention also relates to embodiments and examples obtained by appropriately combining technical means disclosed in different embodiments and examples. Is included in the technical scope.

[1. Microchip)
As shown in FIGS. 1 (a) and 1 (b), the microchip 10 of the present embodiment is a microchip for detecting aggregation of particles (not shown) and contains particles. For the above detection, dry particles are stored which are disposed in the chamber 1 in which light is irradiated to the contained particles and the flow path 5 for flowing the liquid sample into the chamber 1. The storage part 2 is provided.

  In the microchip 10 according to the present embodiment, a storage unit 2 is disposed in a flow path 5 for flowing a liquid sample into the chamber 1, and particles in a dried state are stored in the storage unit 2. ing.

  Therefore, for example, when a liquid sample is injected into the flow channel 5 from the injection unit 6 (as a specific configuration of the injection unit 6, for example, an opening provided at one end of the flow channel 5), the liquid sample is stored. The particles reach the part 2 and dissolve in the liquid sample that reaches the storage part 2.

  Thereafter, the particles dissolved in the liquid sample are accommodated in the chamber 1 while being in contact with the liquid sample. The particles that are in contact with the liquid sample aggregate depending on the components contained in the liquid sample. The movement of the particles from the storage unit 2 to the chamber 1 is performed by the flow of the liquid sample in the flow path.

  In the chamber 1, the particles are irradiated with light, and the aggregation of the particles is detected by detecting transmitted light or scattered light generated as a result of the light irradiation.

  That is, according to the microchip 10 of the present embodiment, by one operation of flowing the liquid sample into the flow path 5, dissolution of particles in the liquid sample, movement of the dissolved particles to the chamber 1, chamber 1 A plurality of steps of irradiating light on the particles inside and detecting transmitted light or scattered light can be performed continuously.

  Below, each structure of the microchip of this Embodiment is demonstrated in detail.

[1-1. particle〕
The microchip of this embodiment is a microchip for detecting particle aggregation. Note that the aggregation of the particles occurs according to the components contained in the liquid sample.

  The specific configuration of the particles is not particularly limited, and can be appropriately selected according to the purpose.

  For example, the particles may be artificially produced particles (for example, metal beads, latex beads, micelles, etc.), or natural particles that exist in nature (for example, dust or dust that becomes an allergen substance, Or a microorganism). Further, the particles may be an aggregate of the same particles or an aggregate of different fine particles.

  A substance capable of binding to a specific component contained in the liquid sample may be bound to the particles. Examples of the substance include, but are not limited to, proteins (eg, antibodies (eg, monoclonal antibodies and polyclonal antibodies)) and nucleic acids (eg, DNA or RNA), but are not limited thereto. Not. As the substance, for example, a substance having a relationship between a ligand and a receptor can be used for a specific component contained in a liquid sample. The substance may be a ligand or the substance may be a receptor.

  If it is the said structure, according to the component contained in a liquid sample, aggregation of particle | grains can be produced efficiently. For example, it is assumed that an antibody is bound to the particle and the antigen of the antibody is contained in the liquid sample. In this case, a plurality of particles are bound to the antigen via the antibody, and as a result, the particles are aggregated. Then, the microchip of the present embodiment detects such particle aggregation.

  As described above, latex beads can be used as the particles. In addition, latex beads to which desired antibodies are bound can be used as particles. According to the said structure, the microchip 10 of this Embodiment can be utilized as a microchip for implementing a latex agglutination immunoturbidimetry.

[1-2. Chamber〕
As shown in FIG. 1A and FIG. 1B, the microchip 10 of the present embodiment includes a chamber 1 in which particles are stored and light is irradiated to the stored particles. .

  The specific configuration of the chamber 1 is not particularly limited as long as the chamber 1 can accommodate particles and can irradiate the particles contained therein with light.

  As shown in FIGS. 3 and 4, the chamber 1 may be formed of, for example, at least a first substrate 20 and a second substrate 21. In this case, the space between the first substrate 20 and the second substrate 21 functions as a space for accommodating the particles 25.

  The shape of each surface (for example, surfaces facing each other) of the first substrate 20 and the second substrate 21 is not particularly limited, and may be a flat surface, a curved surface, or uneven. It may be a flat surface or a curved surface, or any combination thereof.

  The positional relationship between the first substrate 20 and the second substrate 21 is not particularly limited, and the second substrate 21 can be provided at a position where light incident into the chamber 1 through the first substrate 20 can be irradiated. is there.

  For example, it is possible to provide both the first substrate 20 and the second substrate 21 so as to face each other. More specifically, it is possible to provide both of the first substrate 20 and the second substrate 21 so that they face each other in parallel. When both are provided so that the first substrate 20 and the second substrate 21 face each other in parallel, more specifically, the surfaces of the first substrate 20 and the second substrate 21 facing each other are parallel to each other. What should I do. Of course, the present invention is not limited to such an arrangement relationship between the first substrate 20 and the second substrate 21.

  The space inside the chamber 1 can be formed as a space between the first substrate 20 and the second substrate 21. For example, another member (for example, a third substrate and a fourth substrate) is sandwiched between the first substrate 20 and the second substrate 21, and the first substrate 20 and the second substrate 21 are interposed via the another member. It is also possible to form a space inside the chamber 1 by adhering. As said another member, it is possible to use the tape etc. which can be double-sided-bonded. Further, the internal space of the chamber 1 can be formed by integral processing by injection molding or by directly bonding the first substrate 20 and the second substrate 21 with a microchannel pattern.

  Specific configurations of the first substrate 20 and the second substrate 21 are not particularly limited.

  For example, as shown in FIG. 3, it is also possible to use the 1st board | substrate 20 which has a light transmittance, and the 2nd board | substrate 21 which has a light transmittance.

  In this case, light that has entered the chamber 1 through the first substrate 20 passes through the second substrate 21. Therefore, the aggregation of the particles 25 can be detected based on the transmitted light. Of course, since the backscattered light and the forward scattered light are generated from the particles 25 irradiated with the light, the aggregation of the particles 25 can be detected based on the backscattered light and / or the forward scattered light.

  Alternatively, for example, as shown in FIG. 4, the first substrate 20 having optical transparency and at least a part of the light incident into the chamber 1 through the first substrate 20 are reflected toward the inside of the chamber 1. It is also possible to use the second substrate 21.

  In this case, light from a light source (not shown) enters the chamber 1 through the first substrate 20. Then, light immediately after entering the chamber 1 with respect to the particles 25 (in other words, light traveling from the first substrate 20 side to the second substrate 21 side) and the second substrate 21 after entering the chamber 1. (In other words, light traveling from the second substrate 21 side to the first substrate 20 side) is irradiated.

  When the particles 25 are irradiated with light immediately after entering the chamber 1, backscattered light directed toward the first substrate 20 is generated. On the other hand, when the particle 25 is irradiated with the light reflected by the second substrate 21 after entering the chamber 1, forward scattered light directed toward the first substrate 20 is generated.

  That is, according to the above configuration, the back scattered light and the forward scattered light traveling in the same direction can be generated at the same time. Therefore, the detection sensitivity of the scattered light is detected by detecting the scattered light by, for example, one detection unit. In addition, the detection accuracy can be dramatically increased. Therefore, according to the above configuration, the aggregation of the particles 25 can be detected with high accuracy and high sensitivity based on the scattered light (forward scattered light and back scattered light).

  The specific configuration of the light-transmitting first substrate 20 and the light-transmitting second substrate 21 is not particularly limited. For example, at least 50% of the irradiated light, preferably at least 60%, More preferably, the substrate is capable of transmitting at least 70%, more preferably at least 80%, and most preferably at least 90%.

  More specifically, as the first substrate 20 having optical transparency and the second substrate 21 having optical transparency, a glass substrate or an optically transparent resin (for example, PMMA (Poly Methyl Methacrylate), PET ( Poly Ethylene Terephthalate), COC (Cyclo Olefin Copolymer), etc.) can be used.

  The specific configuration of the second substrate 21 that reflects at least part of the light incident into the chamber 1 through the first substrate 20 toward the inside of the chamber 1 is not particularly limited. A specific configuration of the second substrate 21 can be determined based on the light resolution and the absorption capacity of the second substrate 21.

  First, how the light applied to the second substrate 21 is decomposed and absorbed will be described.

  The light irradiated to the second substrate 21 is divided into regular reflection light, scattered light (scattered reflection), and transmitted light, and a part of the light is absorbed by the second substrate 21.

The intensity of light emitted to the second substrate 21 is “I _irradiation ”, the intensity of light reflected by the second substrate 21 is “I _reflection ”, and the intensity of scattered light generated on the second substrate 21 _Is “I _scattering ”, the intensity of light transmitted through the second substrate 21 is “I _transmission ”, and the intensity of light absorbed by the second substrate 21 is “I _absorption ”. Holds. That means
I _irradiation = I _reflection + I _scattering + I _transmission + I _absorption (Relational formula 1).

  The specific configuration of the second substrate 21 can be determined based on the above-described parameters and relational expression 1. An example is shown below.

For example, the second substrate 21 may have a light reflectance that is greater than the light transmittance. In other words, the second substrate 21 may be a substrate that satisfies the condition “I _transmission <I _reflection ”.

Further, the second substrate 21 may have a greater regular reflection light amount than a scattered light amount. In other words, the second substrate 21 may be a substrate that satisfies the condition of “I _scattering <I _reflection ”.

The second substrate 21 has a reflectance of light in the regular reflection direction of 10% or more and less than 100%, and the intensity of light in the regular reflection direction is higher than the intensity of light reflected in other than the regular reflection direction. It can be big. In other words, the second substrate 21 may be a substrate that satisfies the conditions of “0.1 × I _irradiation ≦ I _reflection <1.0 × I _irradiation ” and “I _scattering + I _transmission + I _absorption <I _reflection ”.

  Any of the above configurations can increase the amount of light reflected by the second substrate 21. As a result, the forward scattered light generated as a result of the reflected light being applied to the particles 25 in the chamber 1 can be increased.

  A substrate that satisfies the above-described conditions can be easily selected by measuring each parameter with a known optical device (for example, a spectrophotometer manufactured by Hitachi High-Technologies Corporation). In addition, at the time of filing this application, the data of each parameter described above is well known (see, for example, Chemical Handbook (edited by the Chemical Society of Japan)). Therefore, it is possible to select a substrate that satisfies the above-described conditions based on the document.

  The second substrate 21 may be formed with one configuration or may be formed with a plurality of configurations.

  When the second substrate 21 is formed by a plurality of configurations, for example, the second substrate 21 is formed on the base substrate and the surface of the base substrate that is disposed toward the inside of the chamber 1. And a light reflection layer provided. In the case of this configuration, at least part of the light that enters the chamber 1 through the first substrate 20 is reflected toward the inside of the chamber 1 by the light reflecting layer.

  The structure of the base substrate is not particularly limited, and a desired structure can be used as appropriate. As the base substrate, for example, a light transmissive substrate can be used, and a light non-transmissive substrate can also be used. In the case of the present embodiment, the light reflecting layer is arranged toward the space inside the chamber 1 and light can be efficiently reflected by the light reflecting layer, so that the specific configuration of the base substrate is not particularly limited.

  When the second substrate 21 is formed with a plurality of configurations, for example, the second substrate 21 is formed on the base substrate and the surface of the base substrate that is disposed toward the outside of the chamber. The base substrate may be a light transmissive substrate. In the case of this configuration, at least part of the light that enters the chamber 1 through the first substrate 20 is reflected toward the inside of the chamber 1 by the light reflecting layer.

  In this case, since the light after passing through the base substrate is reflected by the light reflection layer, the base substrate needs to be a light transmissive substrate. The specific configuration of the light transmissive substrate is not particularly limited. For example, a glass substrate or a light transmissive resin (for example, PMMA, PET, COC, or the like) is used in the same manner as the first substrate 20 described above. Is possible.

  In the case where the second substrate 21 is formed with a single configuration, the specific configuration is not particularly limited. For example, a material obtained by processing the material for forming the light reflecting layer into a substrate shape is used as the second substrate. 21 can be used.

  The light reflecting surface of the second substrate 21 (for example, the surface of the second substrate 21 and the light reflecting layer described above) can be formed of a dielectric film (for example, a dielectric multilayer film) and / or a metal film. Although it does not specifically limit as a metal which comprises the said metal film, For example, it may be at least 1 selected from the group which consists of titanium, chromium, copper, aluminum, an aluminum alloy, silver, platinum, and gold | metal | money. Of course, the present invention is not limited to these.

It is also possible to coat a dielectric film as a reflection-enhancing film on the metal film. According to the above configuration, the reflectance of light can be increased. As the material of the dielectric film is not particularly limited, examples thereof _{include TiO 2, Ta 2 O 5,} SiO 2 or MgF _2.

  For example, the second substrate 21 is formed using the above-described metal by a known method (evaporation or sputtering), or at least a part of the surface of the second substrate 21 is coated with the above-described metal. Can be formed.

[1-3. (Flow path)
As shown in FIG. 1A and FIG. 1B, the microchip 10 of the present embodiment is provided with a flow path 5 for flowing a liquid sample into the chamber 1. The chamber 1 is provided at one end of the flow path 5 or in the middle of the flow path 5.

  The number of the flow paths 5 provided for one chamber 1 is not particularly limited, and may be one or plural. If a plurality of flow paths 5 are provided for one chamber 1, continuous measurement can be performed using the microchip 10 of the present embodiment.

  An injection part 6 for injecting a liquid sample into the flow path 5 may be provided at one end of the flow path 5. The injection unit 6 may be any unit that can inject a liquid sample into the channel 5, and the specific configuration is not particularly limited. For example, the injection unit 6 may be an opening or a hole provided at one end of the channel 5. Good.

  An opening or hole different from the injection part 6 may be provided at the other end of the flow path 5. Then, the liquid sample in the chamber 1 may be discharged from the other opening or hole. If both the injection part 6 and an opening or hole different from the injection part 6 are provided, measurement can be continuously performed using the microchip 10 of the present embodiment.

  For example, when a plurality of flow paths 5 are provided in the microchip 10 and a first measurement is performed, a liquid sample is flowed into the chamber 1 using the first flow path 5, and after the measurement, The liquid sample in the chamber 1 is discharged using the flow path 5. Next, when the second measurement is performed, the liquid sample is flowed into the chamber 1 using the second flow path 5, and after the measurement is finished, the liquid in the chamber 1 is used using the second flow path 5. Release the sample. By repeating this, a plurality of measurements can be performed continuously.

  The chamber 1 and the flow path 5 may have the same or different cross-sectional shapes in a direction perpendicular to the liquid feeding direction.

  FIG. 1 (a) shows a microchip 10 when the cross-sectional shape in the direction orthogonal to the liquid feeding direction of the chamber 1 and the flow path 5 (the direction from right to left in the paper surface of FIG. 1 (a)) is different, An example of the situation when viewed from above is shown.

  The microchip 10 shown in FIG. 1A is formed such that the cross-sectional area in the direction perpendicular to the liquid feeding direction of the chamber 1 is larger than the area of the cross section in the direction perpendicular to the liquid feeding direction of the flow path 5. Has been. Although not shown, the microchip 10 is arranged so that the cross-sectional area in the direction perpendicular to the liquid feeding direction of the chamber 1 is smaller than the cross-sectional area in the direction perpendicular to the liquid feeding direction of the flow path 5. It is also possible to form.

  In FIG. 1B, the microchip 10 in the case where the cross-sectional shape in the direction orthogonal to the liquid feeding direction of the chamber 1 and the flow path 5 (the direction from right to left on the paper surface of FIG. 1B) is the same, An example of the state when viewed from above is shown. In this case, a partial region of the flow path 5 functions as the chamber 1.

  According to the above configuration, since the complexity of the configuration of the microchip 10 is reduced, the microchip 10 can be manufactured easily and at low cost.

  In addition, in FIG. 1A and FIG. 1B, the chamber 1 and the flow path 5 are described as separate configurations, but it is also possible to form these configurations as a single configuration. For example, the flow path 5 and the chamber 1 illustrated in FIG. 1B may be configured as a single chamber 1, and the chamber 1 may have both a function as a chamber and a function as a flow path. Is possible. In this case, the entire upper side of the channel 5 and the chamber 1 in FIG. 1B may be the first substrate, and the entire lower side of the channel 5 and the chamber 1 in FIG. 1B may be the second substrate. The same applies to the flow path 5 and the chamber 1 illustrated in FIG.

[1-4. (Storage section)
As shown in FIGS. 1 (a) and 1 (b), the microchip 10 of the present embodiment is provided with a storage unit 2 in which dry particles are stored in a flow path 5. . Specifically, in the microchip 10 of the present embodiment, the storage unit 2 is provided on the upstream side of the chamber 1, in other words, between the injection unit 6 and the chamber 1.

  As described above, in the microchip 10 of the present embodiment, the chamber 1 and the flow path 5 can be formed as a single configuration. In such a case, the storage unit 2 is provided in the flow path 5, in other words, in the chamber 1.

  As used herein, “dried particles” includes a liquid substance (for example, water) only in an amount of 10% by weight or less, more preferably 5% by weight or less, more preferably 1% by weight or less. Only particles, most preferably containing no more than 0.1% by weight. Since “particles” have already been described, description thereof is omitted here.

  The dried particles may contain components other than the particles. Examples of the component include hydrophilic polymers such as polyethylene glycol and saccharides such as sucrose, and one or more of them may be included.

  It is preferable that polyethylene glycol and / or sucrose is contained in the solution containing the particles before drying because the surface of the beads is coated when dried, so that a decrease in activity due to drying can be prevented. I can say that.

  The preservation | save part 2 should just be what can preserve | save the particle | grains of the dried state, and the specific structure is not specifically limited.

  For example, the storage unit 2 is preferably formed of a porous base material. According to the said structure, particle | grains can be preserve | saved with high density in the hole of a porous base material. Moreover, according to the said structure, since the contact area of particle | grains and a liquid sample can be enlarged easily, particle | grains can be efficiently dissolved in a liquid sample.

  The diameter of the holes formed in the porous substrate can be appropriately set according to the diameter of the particles. For example, the diameter of the holes formed in the porous substrate is preferably 5 to 1000 times the diameter of the particle, more preferably 10 to 200 times, and more preferably 50 to 100 times. Most preferably: According to the said structure, particle | grains can be dissolved in a liquid sample more efficiently.

  The size of the storage unit 2 is not particularly limited, but is preferably a size that completely blocks the passage path of the liquid sample in the flow path 5. That is, the size of the storage unit 2 is preferably a size that completely covers the flow path 5 when viewed from the injection unit 6 side.

  According to the above configuration, all liquid samples reach the chamber 1 through the storage unit 2. Therefore, since the particles stored in the storage unit 2 can contact the liquid sample more efficiently, the particles can be more efficiently dissolved in the liquid sample.

[1-5. (Diffusion part)
As shown in FIGS. 2A and 2B, in the microchip 10 of the present embodiment, the inside of the flow path 5 is moved to a position between the storage unit 2 and the chamber 1 in the flow path 5. A diffusing section 3 having a structure for dispersing particles to be dispersed may be provided.

  The diffusing unit 3 may have any structure that disperses particles moving in the flow path 5, and the specific configurations of the diffusing unit 3 and the structure are not particularly limited.

  For example, the structure provided in the diffusing unit 3 may be a plurality of pillar-shaped structures. According to the above configuration, when the particle is moving, the particle collides with the pillar-shaped structure. If the particles collide with the pillar-shaped structure, the moving direction of the particles changes irregularly, and as a result, the particles moving in the flow path 5 can be dispersed.

  The specific shape of the pillar-shaped structure is not particularly limited, and may be, for example, a cone (for example, a cone or a polygonal pyramid) or a column (for example, a cylinder or a polygonal column). .

  The size and density of the structure provided in the diffusing unit 3 are not particularly limited as long as the size and density are such that particles can pass between the structure and the structure.

  For example, the structure provided in the diffusing unit 3 may be a porous substrate. According to the above configuration, the moving direction of the particles changes irregularly as the particles move in the porous substrate, and as a result, the particles moving in the flow path 5 can be dispersed.

The diameter of the holes formed in the porous substrate can be appropriately set according to the diameter of the particles. For example, the diameter of the holes formed in the porous substrate is preferably 10 to 2000 times the diameter of the particles, more preferably 20 to 500 times, and more preferably 100 to 200 times. Most preferably: According to the said structure, particle | grains can be disperse | distributed more efficiently.
[2. Analysis equipment〕
The analyzer according to the present embodiment includes a light source 30 for irradiating light to the chamber 1 of the microchip 10 and transmitted light or scattered light (forward) generated as a result of irradiating the particles in the chamber 1 with light. And a detector 31 for detecting scattered light and / or backscattered light).

  Since the configuration of the microchip 10 has already been described, the description thereof is omitted here.

  More specifically, as shown in FIG. 5A, the analyzer of the present embodiment includes a light source 30 for irradiating light to the chamber 1 of the microchip 10, and a detection surface in the chamber 1. Transmitted light after passing through the chamber 1, which is provided on an extension line in the irradiation direction of light emitted from the light source 30 on the side opposite to the light source 30 (for example, the light irradiation surface in the chamber 1). It is also possible to have a configuration provided with a detection unit 31 for detecting.

  In the case of the analyzer shown in FIG. 5A, the detection unit 31 is a position on an extension line in the irradiation direction of light emitted from the light source 30 and a position where the transmitted light after passing through the chamber 1 can be detected. What is necessary is just to be provided.

  Whether or not the detection unit 31 is provided at the above-described position is determined based on, for example, the light emitted from the light source 30 when the detection unit 31 detects the transmitted light of the microchip 10 that does not contain particles. It can be confirmed by determining whether the strength is 90% or more, more preferably 95% or more, more preferably 99% or more.

  That is, when the transmitted light of the microchip 10 that does not contain particles is detected by the detection unit 31, the detected value is 90% or more, more preferably 95% or more, more preferably 95% or more of the intensity of light emitted from the light source 30. If it is 99% or more, it can be determined that the detection unit 31 is provided at the position described above.

  Note that the analyzer according to the present embodiment may include a determination unit as a configuration for performing the above-described determination.

  Alternatively, as shown in FIG. 5B, the analyzer of the present embodiment includes a light source 30 for irradiating light to the chamber 1 of the microchip 10 and a detection surface (for example, a chamber) in the chamber 1. And a detector 31 for detecting scattered light (forward scattered light and / or back scattered light) provided on the same side as the light source 30 with the light irradiation surface in 1 as a boundary. There may be.

  According to the above configuration, the forward scattered light and the back scattered light generated by the light incident into the chamber 1 can be detected simultaneously by a small number of detectors (for example, one). That is, according to the above configuration, the configuration of the analyzer according to the present embodiment can be simplified.

  In the case of the analyzer shown in FIG. 5B, the detection unit 31 is light (reflected light) in the regular reflection direction by the second substrate of the microchip 10 among the light irradiated from the light source 30 to the microchip 10. ) May not be received. Generally, when the angle between the light beam of the light source 30 and the irradiation surface in the chamber 1 is θ, the position of the detection unit 31 can take an angle other than 180−θ.

  Whether or not the detection unit 31 is provided at the above-described position is determined, for example, when the detection unit 31 detects scattered light from the microchip 10 that does not contain particles, and the detection value is the light emitted from the light source. This can be confirmed by determining whether the strength is 5% or less, more preferably 3% or less, more preferably 1% or less, and most preferably 0%.

  That is, when the scattered light of the microchip 10 that does not contain particles is detected by the detection unit 31, the detected value is 5% or less, more preferably 3% or less, more preferably 3% or less of the intensity of light emitted from the light source. If it is 1% or less, most preferably 0%, it can be determined that the detection unit 31 is provided at the position described above.

  Note that the analyzer according to the present embodiment may include a determination unit as a configuration for performing the above-described determination.

  The specific configuration of the light source 30 is not particularly limited, and a known light source can be used as appropriate. Moreover, the number of the light sources 30 provided in the analyzer of the present embodiment is not particularly limited, and may be one or plural.

  The specific configuration of the detection unit 31 is not particularly limited as long as it can detect transmitted light and / or scattered light. As the detection unit 31, a known detection unit can be used as appropriate. Moreover, the number of the detection parts 31 provided in the analyzer of this Embodiment is not specifically limited, One may be sufficient and plural may be sufficient.

  In the analyzer of the present embodiment, at least one of an optical slit and a condenser lens may be provided in the optical path between the light source 30 and the microchip 10.

  According to the above configuration, since it is possible to irradiate the microchip 10 with light having high intensity, stronger transmitted light and / or scattered light can be generated from the particles. As a result, the detection sensitivity and detection accuracy of transmitted light and / or scattered light can be improved.

  Moreover, according to the said structure, light can be irradiated to the local of the microchip 10. FIG. More specifically, light can be irradiated only on a desired portion of the chamber 1. As a result, it is possible to suppress the generation of reflected light that increases the background during measurement.

  The configuration of the optical slit and the condensing lens is not particularly limited, and a known optical slit and condensing lens can be used as appropriate.

  Although an example of arrangement | positioning of the optical slit 40 (or condensing lens) in the analyzer of this embodiment is shown in FIG. 6, this invention is not limited to this.

  As shown in FIG. 6, in the analyzer of the present embodiment, an optical slit 40 (and / or a condenser lens) is disposed between the light source 30 and the microchip 10 (in other words, the chamber 1). May be. When both the optical slit 40 and the condensing lens are disposed, for example, the optical slit 40 may be disposed near the light source 30 and the condensing lens may be disposed near the microchip 10.

  The light from the light source 30 passes through the optical slit 40 and is then irradiated onto a specific irradiation area 41 of the microchip 10.

  In addition, the light source 30 causes light to enter the chamber 1 among the wall surfaces in which the irradiation area in the direction perpendicular to the liquid feeding direction in the chamber 1 surrounds the internal space of the chamber 1. It is preferable that the first wall surface and light after entering from the first wall surface do not overlap with any other wall surface other than the two wall surfaces of the second wall surface irradiated after traveling straight in the chamber.

  Further, the light source 30 can adjust the length of the irradiation area of the light incident into the chamber 1 in the direction perpendicular to the liquid feeding direction in the chamber 1 in the direction perpendicular to the liquid feeding direction in the chamber 1. It may be a thing.

  More specifically, the light source 30 has a length in a direction perpendicular to the liquid feeding direction in the chamber 1 of an irradiation area of the light incident into the chamber 1 in a direction perpendicular to the liquid feeding direction in the chamber 1. Is adjusted to 50% or more of the length of the chamber 1 in the direction perpendicular to the liquid feeding direction of the chamber 1, more preferably 60% or more, more preferably 70% or more, and further preferably 80% or more. May be.

  Moreover, the light irradiation direction of the light source 30 may be parallel to the liquid feeding direction in the chamber 1 in a direction horizontal to the detection surface in the chamber 1. Specifically, as shown in FIG. 7, the light irradiation direction of the light source 30 is such that the direction A parallel to the detection surface in the chamber 1 is parallel to the liquid feeding direction B in the chamber 1. Also good.

  Further, as shown in FIGS. 8A and 8B, the analyzer 50 of the present embodiment may include an insertion portion 51 for inserting the microchip 10. In addition, the said insertion part 51 may be provided with respect to the analyzer 50, and multiple may be provided.

  It is preferable that the insertion portion 51 can attach and detach the microchip 10 to and from the analyzer 50. According to the said structure, a desired liquid sample can be measured simply and rapidly. In particular, when there are a plurality of liquid samples, the measurement time can be dramatically shortened. Further, according to the above configuration, when the microchip 10 loses desired performance due to deterioration or the like, the microchip 10 can be easily replaced, so that the detection sensitivity and accuracy of measurement can be maintained high. it can.

[Example 1. Preparation of carrier in which dried beads are stored]
0.05% polyethylene glycol and 5% sucrose were added to latex beads (φ0.1 μm) modified with an anti-CRP monoclonal antibody to prepare a bead suspension.

The bead suspension was dropped into a cellulose acetate (ADVANTEC) having a pore size of 3 μm at a volume of 20 μL / cm ² and incubated at 50 ° C. for 30 minutes.

  After 30 minutes, the cellulose acetate was returned to room temperature, and then the cellulose acetate was cut into 2 mm × 2 mm squares with a laser processing machine (GCC), and the cut product was used as a carrier 60 (corresponding to a storage unit). The carrier 60 was stored in a dry state.

[Example 2. Microchip production-1]
9A and 9B show a manufacturing process of the microchip of this embodiment and an outline of the microchip.

  As the chip substrate 61 (corresponding to the second substrate), a Ti thin film (50 nm) -Al (100 nm) thin film (surface is aluminum) substrate in which a thin film was formed on a glass substrate by sputtering was used.

  A double-sided tape 62 (thickness: 50 μm) cut into an I shape (length: 2 cm, width: 600 μm) by a laser processing machine (manufactured by GCC) was attached to the chip substrate 61 described above. Note that the I-shaped cutout portion 64 functions as a flow path.

  Further, a 1.5 cm square glass substrate 63 (corresponding to the first substrate) was bonded to the chip substrate 61 via the double-sided tape 62.

  The carrier 60 described in Example 1 was disposed in the vicinity of one end (corresponding to the injection portion) of the I-shaped cutout portion 64.

[Example 3. Microchip production-2]
An outline of the microchip of this example is shown in FIG.

  As the chip substrate 61 (corresponding to the second substrate), a Ti thin film (50 nm) -Al (100 nm) thin film (surface is aluminum) substrate in which a thin film was formed on a glass substrate by sputtering was used.

  A double-sided tape 62 (thickness: 50 μm) cut into an I shape (length: 2 cm, width: 600 μm) by a laser processing machine (manufactured by GCC) was attached to the chip substrate 61 described above. Note that the I-shaped cutout portion 64 functions as a flow path.

  Further, a 1.5 cm square glass substrate 63 (corresponding to the first substrate) was bonded to the chip substrate 61 via the double-sided tape 62.

  In the vicinity of one end (corresponding to the injection part) of the I-shaped cutout part 64, the carrier 60 described in Example 1 and the filter paper 65 (corresponding to the diffusion part) were arranged.

  As the filter paper 65, qualitative filter paper grade 2 (Whatman) cut into 2 mm × 2 mm by a laser processing machine was used.

[Example 4. Microchip production-3]
An outline of the microchip of this example is shown in FIG.

  As the chip substrate 61 (corresponding to the second substrate), a Ti thin film (50 nm) -Al (100 nm) thin film (surface is aluminum) substrate in which a thin film was formed on a glass substrate by sputtering was used.

As shown in FIG. 11, a microchannel 71 (2 mm × 600 μm × 50 μm) was formed on a PDMS substrate 70 (corresponding to the first substrate) formed by PDMS (POLYDIMETHYLSILOXANE, Toray Dow Corning). A pillar structure 72 was formed in the microchannel 71. In addition, the specific shape of the pillar structure 72 is a cylinder having a lower surface and an upper surface each having a diameter of approximately 5 μm and a height of 50 μm, which is the same as the height of the flow path, and approximately 1 × 10 ⁶ cylinders. It was formed at a density of / cm ² .

  After the chip substrate 61 and the PDMS substrate 70 were bonded together, the carrier 60 described in Example 1 was disposed in the vicinity of one end of the microchannel 71 (corresponding to the injection portion).

[Example 5. Analytical test)
5 μL of a 3.5 μg / dL CRP solution was applied to the microchips described in Examples 2 to 4, and the scattered light was measured after 3 minutes (n = 3).

  At that time, the range of irradiation light was set to 1.9 mm × 1.9 mm by using pinholes.

[Example 6. result of analysis〕
In any of the microchips of Examples 2 to 4, aggregation of latex beads was observed. The results are shown in FIG.

  In the microchip of Example 2, the CV value was 9.17%, but in the microchips of Example 3 and Example 4, the CV value could be suppressed to 5.52% and 4.71%, respectively. It was.

  The present invention is not limited to the configurations described above, and various modifications can be made within the scope of the claims, and technical means disclosed in different embodiments and examples are appropriately used. Embodiments and examples obtained in combination are also included in the technical scope of the present invention.

  The present invention can be used for a microchip for performing various measurement methods (for example, latex agglutination immunoturbidimetry, turbidimeter, particle counter, etc.). More specifically, the present invention can be used for a turbidimeter or a particle counter.

1: Chamber 2: Storage unit 3: Diffusion unit 5: Channel 6: Injection unit 10: Microchip 20: First substrate 21: Second substrate 25: Particles 30: Light source 31: Detection unit 40: Optical slit 41: Irradiation Area 50: Analyzer 51: Insertion unit 60: Carrier (storage unit)
61: Chip substrate (second substrate)
62: Double-sided tape 63: Glass substrate (first substrate)
64: I-shaped cutout (flow path)
65: Filter paper (diffusion part)
70: PDMS substrate (first substrate)
71: Microchannel (flow path)
72: Pillar structure

Claims (15)

A microchip for detecting particle aggregation,
A chamber in which the particles are contained and light is applied to the particles for the detection;
A microchip, comprising: a storage unit in which the particles in a dried state are stored, which is disposed in a flow path for flowing a liquid sample into the chamber.   The microchip according to claim 1, wherein the storage unit is formed of a porous substrate.   2. A diffusion unit including a structure for dispersing the particles moving in the flow channel is provided at a position between the storage unit and the chamber in the flow channel. Or the microchip of 2.   The microchip according to claim 3, wherein the structure is a plurality of pillar-shaped structures.   The microchip according to claim 3, wherein the structure is a porous substrate.   6. The microchip according to any one of claims 1 to 5, wherein the flow path is provided with an injection part for injecting a liquid sample into the flow path.   The microchip according to any one of claims 1 to 6, wherein the particles are latex particles or latex particles to which an antibody is bound.   The chamber reflects at least a part of the light that enters the chamber through the first substrate and at least a first substrate having light transmissivity to which the light is irradiated, toward the inside of the chamber. The microchip according to claim 1, wherein the microchip is formed by a second substrate. A light source for irradiating light to the chamber of the microchip according to any one of claims 1 to 7,
For detecting the transmitted light after passing through the chamber, provided on the extension line in the irradiation direction of the light emitted from the light source on the opposite side of the light source with the detection surface in the chamber as a boundary And an analysis device. A light source for irradiating light to the chamber of the microchip according to any one of claims 1 to 8,
And a detection unit for detecting scattered light, which is provided on the same side as the light source with a detection surface in the chamber as a boundary.   The analyzer according to claim 9 or 10, wherein at least one of an optical slit and a condenser lens is provided in an optical path between the light source and the microchip.   The light source is a first unit that causes light to enter the chamber out of a wall surface in which an irradiation area in a direction orthogonal to the liquid feeding direction in the chamber surrounds the internal space of the chamber. The wall surface and the light after entering from the first wall surface do not overlap with any wall surface other than the two wall surfaces of the second wall surface irradiated after traveling straight through the chamber. The analyzer according to any one of 11.   The light source is capable of adjusting the length of the irradiation area in the direction perpendicular to the liquid feeding direction in the chamber to the direction perpendicular to the liquid feeding direction in the chamber. The analyzer according to any one of claims 9 to 12, wherein the analyzer is provided.   The light source has a length in a direction perpendicular to the liquid feeding direction in the chamber of an irradiation area in a direction perpendicular to the liquid feeding direction in the chamber. The analyzer according to claim 13, wherein the analyzer is adjusted to 80% or more of the length of the chamber in a direction perpendicular to the liquid direction.   15. The light irradiation direction of the light source is such that a direction horizontal to the detection surface in the chamber is parallel to a liquid feeding direction in the chamber. The analyzer described.

Images